Prange-Kiel J, Rune G M, Zwirner M, Wallwiener D, Kiesel L
Institut of Anatomy, Ernst-Moritz-Arndt University, Greifswald, Germany.
Exp Clin Endocrinol Diabetes. 2001;109(4):231-7. doi: 10.1055/s-2001-15111.
The effects of RU 486 together with estradiol and progesterone on estrogen receptor alpha and progesterone receptor (isoforms A and B) expression were studied in human endometrial long term cultures at the mRNA and protein level. We asked whether ligand induced receptor regulation, found in mammals in vivo, is also found in human cultured endometrial cells with special regard to the progesterone isoforms A and B. Endometrial cultures were maintained for 27 days. Media were supplemented with progesterone and/or estradiol alone or in combination with RU 486. Receptor expression (estrogen receptor alpha and progesterone receptor isoform A and B) was examined at the mRNA level by RT-PCR and at the protein level by western blot analysis. All receptor types examined were expressed in our culture model. Estradiol led to a general increase of receptor expression whereas treatment with estradiol in combination with progesterone down regulated receptor expression. The receptor down regulation was not found when RU 486 was additionally supplemented into the medium. Activation or inhibition of expression due to these treatments was similar for both PR isoforms. Our results (1) show that in our culture system estradiol induced up regulation of estrogen receptor and progesterone receptor A and B and suggest that the estrogen induced up regulation is prevented by progesterone (2) a clear cut antigestagenic effect of RU 486 and (3) suggest that both progesterone isoforms are analogously regulated in our culture model. We conclude that human endometrial cell cultures are suitable for the study of the dynamics of steroid receptor expression.
在人子宫内膜长期培养物中，研究了RU 486联合雌二醇和孕酮对雌激素受体α及孕酮受体（A和B亚型）表达的影响，检测水平为mRNA和蛋白质水平。我们探讨了在哺乳动物体内发现的配体诱导的受体调节现象，在人培养的子宫内膜细胞中，尤其是孕酮A和B亚型中是否也存在。子宫内膜培养物维持27天。培养基单独添加孕酮和/或雌二醇，或与RU 486联合添加。通过RT-PCR检测mRNA水平的受体表达（雌激素受体α及孕酮受体A和B亚型），通过蛋白质印迹分析检测蛋白质水平的受体表达。所有检测的受体类型在我们的培养模型中均有表达。雌二醇导致受体表达普遍增加，而雌二醇与孕酮联合处理则下调受体表达。当培养基中额外添加RU 486时，未发现受体下调现象。两种孕酮亚型因这些处理导致的表达激活或抑制相似。我们的结果：（1）表明在我们的培养系统中，雌二醇诱导雌激素受体以及孕酮受体A和B上调，提示雌激素诱导的上调被孕酮阻止；（2）RU 486有明确的抗孕激素作用；（3）提示在我们的培养模型中，两种孕酮亚型受到类似调节。我们得出结论，人子宫内膜细胞培养物适用于研究类固醇受体表达的动态变化。
